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cAMP-Glo™ Assay

The cAMP-Glo™ Assay is a homogeneous, bioluminescent and high-throughput assay for measuring cAMP levels in cells. The cAMP-Glo™ Assay monitors cAMP production in cells in response to the effects of test compounds on G protein-coupled receptors (GPCR). GPCRs that couple with adenylate cyclase will increase or decrease intracellular cAMP. The assay is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction.

The cAMP-Glo™ Assay is a homogeneous, bioluminescent and high-throughput assay for measuring cAMP levels in cells. The cAMP-Glo™ Assay monitors cAMP production in cells in response to the effects of test compounds on G protein-coupled receptors (GPCR). GPCRs that couple with adenylate cyclase will increase or decrease intracellular cAMP. The assay is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction.

The cAMP-Glo™ Assay can be performed in 96-, 384- or 1536-well plates. The cells are induced with a test compound for an appropriate period of time to modulate cAMP levels. After induction, cells are lysed to release cAMP, then the cAMP detection solution, which contains protein kinase A, is added. The Kinase-Glo® Reagent is then added to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. Luminescence can be correlated to the cAMP concentrations by using a cAMP standard curve. The half-life for the luminescent signal is greater than 4 hours. This extended signal half-life eliminates the need for luminometers with reagent injectors and allows batch-mode processing of multiple plates.

Features - Benefits

Fast and Easy to Use:Assay can be completed in approximately 45 minutes. Homogeneous. Two steps following lysis of cells.

Applications

Notes

Cat.# V1501 contains sufficient reagents to perform 300 assays at 60μl final volume per well in 384-well plates. Cat.# V1502 contains sufficient reagents to perform 3,000 assays at 60μl final volume per well in 384-well plates. Cat.# V1503 contains sufficient reagents to perform 30,000 assays at 60μl final volume per well in 384-well plates.

This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.

Storage Conditions

Store the system at –20°C. Once prepared, the cAMP detection solution (cAMP-Glo™ Reaction Buffer with Protein Kinase A) should not be frozen. Once prepared, the Kinase-Glo® Reagent should be dispensed into aliquots and stored at –20°C. See the product label for the expiration date.

For product intended use please see Patents & Disclaimers tab.

Figure 1. Representation of cAMP production in cells and the cAMP-Glo™ Assay.

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